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EP0299028B1 - Process for producing l-aminoacids - Google Patents

Process for producing l-aminoacids Download PDF

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Publication number
EP0299028B1
EP0299028B1 EP88901022A EP88901022A EP0299028B1 EP 0299028 B1 EP0299028 B1 EP 0299028B1 EP 88901022 A EP88901022 A EP 88901022A EP 88901022 A EP88901022 A EP 88901022A EP 0299028 B1 EP0299028 B1 EP 0299028B1
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group
groups
formula
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general formula
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EP0299028A1 (en
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Uwe Klages
Alfred Weber
Ludwig Wilschowitz
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Bayer Pharma AG
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Schering AG
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P13/00—Preparation of nitrogen-containing organic compounds
    • C12P13/04—Alpha- or beta- amino acids
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S435/00—Chemistry: molecular biology and microbiology
    • Y10S435/8215—Microorganisms
    • Y10S435/822—Microorganisms using bacteria or actinomycetales
    • Y10S435/872—Nocardia

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  • the invention relates to a process for the preparation of L-amino acids of the general formula I.
  • R 1 is an alkyl radical with a maximum of 12 carbon atoms which is optionally substituted by hydroxyl groups, mercapto groups, halogen atoms, amino groups, carbonyl groups or guanidino groups and / or interrupted by oxygen atoms, nitrogen atoms or sulfur atoms, and in the case of the mercapto compounds of the formula I also their dithio compounds, which is characterized in that that the microorganism Nocardia spec. DSM 3306 on a D, L-imidazolidinedione derivative of the general formula II wherein R1 has the meaning given above or, in the case of the mercapto compounds of the formula II, can also act on their dithio compounds.
  • the D, L-imidazolidinedione derivatives of the general formula II and consequently also the L-amino acids of the general formula I prepared therefrom can, for example, be the methyl group, the ethyl group, the propyl group, carry the 1-methylethyl group, the butyl group, the 1-methylpropyl group, the 2-methylpropyl group, the 1,1-dimethylethyl group, the pentyl group, the 1-methylbutyl group, the 3-methylbutyl group or the hexyl group.
  • Particularly preferred alkyl groups R1 are those with a maximum of 6 carbon atoms, such as the methyl group of the process product alanine, the 1-methylethyl group of valine, the 2-methylpropyl group of leucine, the 1-methylpropyl group of isoleucine or the ethyl group of ⁇ -aminobutyric acid.
  • the alkyl groups R1 can optionally be substituted by hydroxyl groups, mercapto groups, halogen atoms, amino groups, carbonyl groups or guanidino groups, monosubstituted alkyl groups being preferred, or interrupted by oxygen atoms (preferably one), nitrogen atoms (preferably one or two) or sulfur atoms (preferably one) .
  • alkyl groups which are substituted by hydroxyl groups or mercapto groups the hydroxymethyl group of serine, the 1-hydroxyethyl group of threonine, the mercaptomethyl group of cysteine, the 2-mercaptoethyl group of homocysteine and the 1-mercapto-1-methyl-ethyl group of ⁇ are particularly emphasized -Thiovalins.
  • the 2-methylthioethyl group of methionine should be emphasized.
  • Alkyl groups R1, which carry an amino group or a guanidino group as substituents R1 are, for example, the 2-aminoethyl group of ⁇ , ⁇ -diaminobutyric acid, the 3-aminopropyl group of ornithine, the 3-guanidinopropyl group of arginine and the 4-aminobutyl group of lysine.
  • Suitable oxoalkyl groups R 1 are preferably those which are additionally substituted by a hydroxyl group, amino group or guanidino group and / or interrupted by an oxygen atom or a nitrogen atom - more precisely an imino group; such groups are, for example, the acetoxymethyl group of O-acetylserine, the 1-acetoxyethyl group of 0-acetylthreonine, the carboxymethyl group of aspartic acid, the 2-carboxyethyl group of glutamic acid, the 2-methoxy-2-oxoethyl group of ⁇ -aspartic acid monomethyl ester, the 3-ureidopropyl group Citrulins or the 3-guanidino-3-oxo-propyl group of canavanine.
  • the process according to the invention is carried out using the microorganism Nocardia spec. DSM 3306 carried out.
  • This microorganism was isolated from soil samples by adding a mineral salt medium containing 5- (2-methylpropyl) hydantoin as the sole nitrogen source, incubating them and growing the cultures on agar plates which were also 5- (2-methylpropyl) hydantoin as the only source of nitrogen contained.
  • microorganism obtained was deposited with the German Collection for Microorganisms and received the number DSM 3306 there. It is irrevocably available to experts.
  • Taxonomically it has the following properties: Colony morphology round, irregular edge, not translucent, Cell morphology in young cultures, rods 1.2 ⁇ m thick, 8-20 ⁇ m long, fragment to rods 2-5 ⁇ m long, unbranched, immobile Gram stain gram positive Acid resistance negative Endospores negative Oxygen ratio obligatory aerobic Catalase positive Oxidase positive Optimal temperature 30-37 ° C Citrate utilization negative Nitrite from nitrate positive Indole formation negative Methyl red negative Voges-Proskauer negative Urease negative H2S education negative Gelatin liquefaction positive Starch hydrolysis positive Sugar utilization Acid from sucrose, glucose, fructose, arabinose no gas formation NaCl tolerance until 5%.
  • This microorganism is grown under the culture conditions usually used in a suitable nutrient medium with aeration, submerged cultures. Then the substrate (preferably dissolved in a suitable solvent) is added to the culture and fermented until a maximum substrate conversion is reached.
  • Suitable substrate solvents are, for example, water, methanol, ethanol, glycol monomethyl ether, dimethylformamide or dimethyl sulfoxide.
  • the optimal substrate concentration, substrate addition time and fermentation time depend on the structure of the substrate used and the type of fermentation conditions used. As is generally necessary in the case of microbiological steroid conversions, these quantities must be determined in individual cases by preliminary tests as are known to the person skilled in the art.
  • the pH of the fermentation broth is preferably adjusted to a pH of 7.5-10.
  • the cultured microorganism from the culture medium, for example by filtration or centrifugation, to immobilize it if desired using one of the known methods and to carry out the fermentation of the substrates with the isolated cell mass using the resting cell method or by means of the immobilizates.
  • a 500 ml Erlenmeyer flask with 100 ml sterile culture medium containing 0.5 g meat extract, 0.5 g peptone, 0.5 g yeast extract and 0.2 g sodium chloride is mixed with Nocardia spec. Inoculated DSM 3306 and shaken for 20 hours at 30 ° C at 180 revolutions / minute. Then the cell mass is separated by centrifugation and washed with physiological saline.
  • Example 2 Under the conditions of Example 1, 3.5 mg of L-isoleucine are formed from 10 mg of 5- (1-methylpropyl) hydantoin.
  • a 2 l Erlenmeyer flask with 500 ml sterile culture medium containing 2.5 g meat extract, 2.5 g peptone, 2.5 g yeast extract and 1 g of sodium chloride is mixed with 50 ml of a 20 hour old culture of Nocardia spec.
  • L-valine 2.1 mg are formed from 10 mg of 5- (1-methyl-ethyl) -hydantoin under the conditions of Example 6b.
  • cystine are formed from 10 mg of 5,5'-dithiobismethylene-bis-hydantoin under the conditions of Example 6b.

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Abstract

PCT No. PCT/DE88/00037 Sec. 371 Date Sep. 23, 1988 Sec. 102(e) Date Sep. 23, 1988 PCT Filed Jan. 22, 1988 PCT Pub. No. WO88/05468 PCT Pub. Date Jul. 28, 1988.Process is described for the production of L-amino acids of general formula I <IMAGE> (I) in which R1 means an alkyl radical with at most 12 carbon atoms optionally substituted by hydroxy groups, mercapto groups, halogen atoms, amino groups, carbonyl groups or guanidino groups and/or interrupted by oxygen atoms, nitrogen atoms or sulfur atoms, and in the case of mercapto compounds of formula I also their dithio compounds, characterized in that the microorganism Nocardia spec. DSM 3306 or its enzymes are allowed to act on a D,L-imidazolidinedione derivative of general formula II <IMAGE> (II) in which R1 has the above-named meaning or, in the case of mercapto compounds of formula II, also in their dithio compounds.

Description

Die Erfindung betrifft ein Verfahren zur Herstellung von L-Aminosäuren der allgemeinen Formel I

Figure imgb0001

worin
R₁ einen gegebenenfalls durch Hydroxygruppen, Mercaptogruppen, Halogenatome, Aminogruppen, Carbonylgruppen oder Guanidinogruppen substituierter und/oder durch Sauerstoffatome, Stickstoffatome oder Schwefelatome unterbrochener Alkylrest mit maximal 12 Kohlenstoffatomen bedeutet, und im Falle der Mercaptoverbindungen der Formel I auch deren Dithioverbindungen, welches dadurch gekennzeichnet ist, daß man den Mikroorganismus Nocardia spec. DSM 3306 auf ein D,L-Imidazolidindion-Derivat der allgemeinen Formel II
Figure imgb0002
worin
R₁ die obengenannte Bedeutung besitzt oder im Falle der Mercaptoverbindungen der Formel II auch auf deren Dithioverbindungen einwirken läßt.The invention relates to a process for the preparation of L-amino acids of the general formula I.
Figure imgb0001
wherein
R 1 is an alkyl radical with a maximum of 12 carbon atoms which is optionally substituted by hydroxyl groups, mercapto groups, halogen atoms, amino groups, carbonyl groups or guanidino groups and / or interrupted by oxygen atoms, nitrogen atoms or sulfur atoms, and in the case of the mercapto compounds of the formula I also their dithio compounds, which is characterized in that that the microorganism Nocardia spec. DSM 3306 on a D, L-imidazolidinedione derivative of the general formula II
Figure imgb0002
wherein
R₁ has the meaning given above or, in the case of the mercapto compounds of the formula II, can also act on their dithio compounds.

Die D,L-Imidazolidindion-Derivate der allgemeinen Formel II und demzufolge auch die daraus hergestellten L-Aminosäuren der allgemeinen Formel I können als Substituenten R₁ beispielsweise die Methylgruppe, die Ethylgruppe, die Propylgruppe, die 1-Methylethylgruppe, die Butylgruppe, die 1-Methylpropylgruppe, die 2-Methylpropylgruppe, die 1,1-Dimethylethylgruppe, die Pentylgruppe, die 1-Methylbutylgruppe, die 3-Methylbutylgruppe oder die Hexylgruppe tragen. Besonders bevorzugte Alkylgruppen R₁ sind solche mit maximal 6 Kohlenstoffatomen, wie die Methylgruppe des Verfahrensprodukts Alanin, die 1-Methylethylgruppe des Valins, die 2-Methylpropylgruppe des leucins, die 1-Methylpropylgruppe des Isoleucins oder die Ethylgruppe der α-Aminobuttersäure.The D, L-imidazolidinedione derivatives of the general formula II and consequently also the L-amino acids of the general formula I prepared therefrom can, for example, be the methyl group, the ethyl group, the propyl group, carry the 1-methylethyl group, the butyl group, the 1-methylpropyl group, the 2-methylpropyl group, the 1,1-dimethylethyl group, the pentyl group, the 1-methylbutyl group, the 3-methylbutyl group or the hexyl group. Particularly preferred alkyl groups R₁ are those with a maximum of 6 carbon atoms, such as the methyl group of the process product alanine, the 1-methylethyl group of valine, the 2-methylpropyl group of leucine, the 1-methylpropyl group of isoleucine or the ethyl group of α-aminobutyric acid.

Die Alkylgruppen R₁ können gegebenenfalls durch Hydroxygruppen, Mercaptogruppen, Halogenatome, Aminogruppen, Carbonylgruppen oder Guanidinogruppen substituiert sein, wobei einfach substituierte Alkylgruppen bevorzugt sind, oder durch Sauerstoffatome (vorzugsweise eins), Stickstoffatome (vorzugsweise eins oder zwei) oder Schwefelatome (vorzugsweise eins) unterbrochen sein. Als Alkylgruppen, die durch Hydroxygruppen oder Mercaptogruppen substituiert sind, seien besonders hervorgehoben die Hydroxymethylgruppe des Serins, die 1-Hydroxyethylgruppe des Threonins, die Mercaptomethylgruppe des Cysteins, die 2-Mercaptoethylgruppe des Homocysteins und die 1-Mercapto-1-methyl-ethylgruppe des β-Thiovalins. Als eine durch ein Schwefelatom unterbrochene Alkylgruppe R₁ sei die 2-Methylthioethylgruppe des Methionins hervorgehoben. Alkylgruppen R₁, die als Substituenten R₁ eine Aminogruppe oder eine Guanidinogruppe tragen sind beispielsweise die 2-Aminoethylgruppe der α,γ-Diaminobuttersäure, die 3-Aminopropylgruppe des Ornithins, die 3-Guanidinopropylgruppe des Arginins und die 4-Aminobutylgruppe des lysins. Als Oxoalkylgruppen R₁ kommen vorzugsweise solche in Betracht, die zusätzlich noch durch eine Hydroxygruppe, Aminogruppe oder Guanidinogruppe substituiert und/oder durch ein Sauerstoffatom oder ein Stickstoffatom - genauer eine Iminogruppe - unterbrochen sind; derartige Gruppen sind beispielsweise die Acetoxymethylgruppe des O-Acetylserins, die 1-Acetoxyethylgruppe des 0-Acetylthreonins, die Carboxymethylgruppe der Asparaginsäure, die 2-Carboxyethylgruppe der Glutaminsäure, die 2-Methoxy-2-oxoethylgruppe des ω-Asparaginsäuremonomethylesters, die 3-Ureidopropylgruppe des Citrulins oder die 3-Guanidino-3-oxo-propylgruppe des Canavanins.The alkyl groups R₁ can optionally be substituted by hydroxyl groups, mercapto groups, halogen atoms, amino groups, carbonyl groups or guanidino groups, monosubstituted alkyl groups being preferred, or interrupted by oxygen atoms (preferably one), nitrogen atoms (preferably one or two) or sulfur atoms (preferably one) . As alkyl groups which are substituted by hydroxyl groups or mercapto groups, the hydroxymethyl group of serine, the 1-hydroxyethyl group of threonine, the mercaptomethyl group of cysteine, the 2-mercaptoethyl group of homocysteine and the 1-mercapto-1-methyl-ethyl group of β are particularly emphasized -Thiovalins. As an alkyl group interrupted by a sulfur atom R₁, the 2-methylthioethyl group of methionine should be emphasized. Alkyl groups R₁, which carry an amino group or a guanidino group as substituents R₁ are, for example, the 2-aminoethyl group of α, γ-diaminobutyric acid, the 3-aminopropyl group of ornithine, the 3-guanidinopropyl group of arginine and the 4-aminobutyl group of lysine. Suitable oxoalkyl groups R 1 are preferably those which are additionally substituted by a hydroxyl group, amino group or guanidino group and / or interrupted by an oxygen atom or a nitrogen atom - more precisely an imino group; such groups are, for example, the acetoxymethyl group of O-acetylserine, the 1-acetoxyethyl group of 0-acetylthreonine, the carboxymethyl group of aspartic acid, the 2-carboxyethyl group of glutamic acid, the 2-methoxy-2-oxoethyl group of ω-aspartic acid monomethyl ester, the 3-ureidopropyl group Citrulins or the 3-guanidino-3-oxo-propyl group of canavanine.

Das erfindungsgemäße Verfahren wird unter Verwendung des Mikroorganismus Nocardia spec. DSM 3306 durchgeführt. Dieser Mikroorganismus wurde aus Erdproben isoliert, indem man diese mit einem Mineralsalzmedium, welches 5-(2-Methylpropyl)-hydantoin als einzige Stickstoffquelle enthielt, versetzte, incubierte und die gewachsenen Kulturen auf Agarplatten, die ebenfalls 5-(2-Methylpropyl)-hydantoin als einzige Stickstoffquelle enthielten, ausplattete.The process according to the invention is carried out using the microorganism Nocardia spec. DSM 3306 carried out. This microorganism was isolated from soil samples by adding a mineral salt medium containing 5- (2-methylpropyl) hydantoin as the sole nitrogen source, incubating them and growing the cultures on agar plates which were also 5- (2-methylpropyl) hydantoin as the only source of nitrogen contained.

Der erhaltene Mikroorganismus wurde bei der Deutschen Sammlung für Mikroorganismen hinterlegt und erhielt dort die Nummer DSM 3306. Er steht der Fachwelt unwiderruflich zur Verfügung.The microorganism obtained was deposited with the German Collection for Microorganisms and received the number DSM 3306 there. It is irrevocably available to experts.

Er besitzt taxonomisch folgende Eigenschaften: Koloniemorphologie rund, unregelmäßiger Rand, nicht durchscheinend, Zellmorphologie in jungen Kulturen Stäbchen 1,2 µm dick, 8-20 µm lang, fragmentieren zu Stäbchen von 2-5 µm Länge, unverzweigt, unbeweglich Gram-Färbung gram-positiv Säurefestigkeit negativ Endosporen negativ Sauerstoffverhältnis obligat aerob Katalase positiv Oxidase positiv Temperaturoptimum 30-37°C Citratverwertung negativ Nitrit aus Nitrat positiv Indolbildung negativ Methylrot negativ Voges-Proskauer negativ Urease negativ H₂S-Bildung negativ Gelatineverflüssigung positiv Stärkehydrolyse positiv Zuckerverwertung Säure aus Saccharose, Glucose, Fructose, Arabinose keine Gasbildung NaCl-Toleranz bis 5%. Taxonomically, it has the following properties: Colony morphology round, irregular edge, not translucent, Cell morphology in young cultures, rods 1.2 µm thick, 8-20 µm long, fragment to rods 2-5 µm long, unbranched, immobile Gram stain gram positive Acid resistance negative Endospores negative Oxygen ratio obligatory aerobic Catalase positive Oxidase positive Optimal temperature 30-37 ° C Citrate utilization negative Nitrite from nitrate positive Indole formation negative Methyl red negative Voges-Proskauer negative Urease negative H₂S education negative Gelatin liquefaction positive Starch hydrolysis positive Sugar utilization Acid from sucrose, glucose, fructose, arabinose no gas formation NaCl tolerance until 5%.

Aufgrund seiner morphologischen und physiologischen Eigenschaften wurde der Stamm nach "Bergey's Manual of Determinative Bacteriology", 8. Auflage (1974), vorläufig der Gattung Nocardia zugeordnet.Because of its morphological and physiological properties, the strain was provisionally assigned to the genus Nocardia according to "Bergey's Manual of Determinative Bacteriology", 8th edition (1974).

Dieser Mikroorganismus wird unter den üblicherweise verwendeten Kulturbedingungen in einem geeigneten Nährmedium unter Belüften, Submerskulturen angezüchtet. Dann setzt man der Kultur das Substrat (vorzugsweise in einem geeigneten Lösungsmittel gelöst) zu und fermentiert, bis eine maximale Substratumwandlung erreicht ist.This microorganism is grown under the culture conditions usually used in a suitable nutrient medium with aeration, submerged cultures. Then the substrate (preferably dissolved in a suitable solvent) is added to the culture and fermented until a maximum substrate conversion is reached.

Geeignete Substratlösungsmittel sind beispielsweise Wasser, Methanol, Äthanol, Glykolmonomethyläther, Dimethylformamid oder Dimethylsulfoxyd.Suitable substrate solvents are, for example, water, methanol, ethanol, glycol monomethyl ether, dimethylformamide or dimethyl sulfoxide.

Die optimale Substratkonzentration, Substratzugabezeit und Fermentationsdauer ist von der Struktur des verwendeten Substrates und der Art der verwendeten Fermentationsbedingungen abhängig. Diese Größen müssen, wie dies bei mikrobiologischen Steroidumwandlungen allgemein erforderlich ist, im Einzelfall durch Vorversuche, wie sie dem Fachmann geläufig sind, ermittelt werden. Während der Fermentation wird der pH-Wert der fermentationsbrühe vorzugsweise auf einen pH-Wert von 7,5-10 eingestellt.The optimal substrate concentration, substrate addition time and fermentation time depend on the structure of the substrate used and the type of fermentation conditions used. As is generally necessary in the case of microbiological steroid conversions, these quantities must be determined in individual cases by preliminary tests as are known to the person skilled in the art. During the fermentation, the pH of the fermentation broth is preferably adjusted to a pH of 7.5-10.

Andererseits ist es aber auch möglich, den angezüchteten Mikroorganismus beispielsweise durch Filtration oder Zentrifugieren vom Kulturmedium abzutrennen, ihn gewünschtenfalls mittels einer der bekannten Methoden zu immobilisieren und die Fermentation der Substrate mit der isolierten Zellmasse im resting cell Verfahren oder mittels der Immobilisate durchzuführen.On the other hand, it is also possible to separate the cultured microorganism from the culture medium, for example by filtration or centrifugation, to immobilize it if desired using one of the known methods and to carry out the fermentation of the substrates with the isolated cell mass using the resting cell method or by means of the immobilizates.

Die nachfolgenden Ausführungsbeispiele dienen zur Erläuterung des erfindungsgemäßen Verfahrens.The following exemplary embodiments serve to explain the method according to the invention.

Beispiel 1example 1

Ein 500 ml Erlenmeyerkolben mit 100 ml sterilem Nährmedium enthaltend 0,5 g Fleischextrakt, 0,5 g Pepton, 0,5 g Hefeextrakt und 0,2 g Natriumchlorid wird mit Nocardia spec. DSM 3306 beimpft und 20 Stunden lang bei 30° C mit 180 Umdrehungen/Minute geschüttelt. Dann trennt man die Zellmasse durch Zentrifugieren ab und wäscht sie mit physiologischer Kochsalzlösung.A 500 ml Erlenmeyer flask with 100 ml sterile culture medium containing 0.5 g meat extract, 0.5 g peptone, 0.5 g yeast extract and 0.2 g sodium chloride is mixed with Nocardia spec. Inoculated DSM 3306 and shaken for 20 hours at 30 ° C at 180 revolutions / minute. Then the cell mass is separated by centrifugation and washed with physiological saline.

400 mg der feuchten Zellmasse werden in 10 ml 0,1 M Tris/HCl-Puffer vom pH 8,5 suspendiert, mit 10 mg 5-(2-Methylpropyl)-hydantoin versetzt und 24 Stunden lang bei 30° C incubiert. Durch Bestimmung mit L-Aminosäure-Oxidase wird ermittelt, daß 7,9 mg L-Leucin gebildet werden.400 mg of the moist cell mass are suspended in 10 ml of 0.1 M Tris / HCl buffer with a pH of 8.5, 10 mg of 5- (2-methylpropyl) hydantoin are added and the mixture is incubated at 30 ° C. for 24 hours. By determination with L-amino acid oxidase it is determined that 7.9 mg L-leucine are formed.

Beispiel 2Example 2

Unter den Bedingungen des Beispiels 1 werden aus 10 mg 5-(1-Methylpropyl)-hydantoin 3,5 mg L-Isoleucin gebildet.Under the conditions of Example 1, 3.5 mg of L-isoleucine are formed from 10 mg of 5- (1-methylpropyl) hydantoin.

Beispiel 3Example 3

Unter den Bedingungen des Beispiels 1 werden aus 10 mg 5-(1-Methylethyl)-hydantoin 2,5 mg L-Valin gebildet.Under the conditions of Example 1, 2.5 mg of L-valine are formed from 10 mg of 5- (1-methylethyl) hydantoin.

Beispiel 4Example 4

Unter den Bedingungen des Beispiels 1 werden aus 10 mg 5,5'-Dithiobismethylen-bis-hydantoin 2,1 mg Cystin gebildet.Under the conditions of Example 1, 10 mg of 5,5'-dithiobismethylene-bis-hydantoin form 2.1 mg of cystine.

Beispiel 5Example 5

Ein 2 l Erlenmeyerkolben mit 500 ml sterilem Nährmedium enthaltend 2,5 g Fleischextrakt, 2,5 g Pepton, 2,5 g Hefeextrakt und 1 g Natriumchlorid wird mit 50 ml einer 20 Stunden alten Kultur von Nocardia spec. DSM 3306 - hergestellt nach Beispiel 1 - beimpft und 20 Stunden lang bei 30° C mit 180 Umdrehungen pro Minute geschüttelt. Die Zellmasse wird abzentrifugiert und mit physiologischer Kochsalzlösung gewaschen.A 2 l Erlenmeyer flask with 500 ml sterile culture medium containing 2.5 g meat extract, 2.5 g peptone, 2.5 g yeast extract and 1 g of sodium chloride is mixed with 50 ml of a 20 hour old culture of Nocardia spec. DSM 3306 - manufactured according to Example 1 - inoculated and shaken for 20 hours at 30 ° C at 180 revolutions per minute. The cell mass is centrifuged off and washed with physiological saline.

8 g der feuchten Zellmasse werden in 200 ml 0,1 M Tris/HCl-Puffer vom pH 8,5 suspendiert mit 1,0 g 5-(2-Methylpropyl)-hydantoin versetzt und 24 Stunden lang bei 30° C incubiert. Man zentrifugiert die Zellmasse dann ab und isoliert aus dem Filtrat 660 mg L-Leucin vom Zersetzungspunkt 291° C (aus wäßrigem Ethanol) [α] 20 D 

Figure imgb0003
= + 15,6° (20 %ige wäßrige Salzsäure).8 g of the moist cell mass are suspended in 200 ml of 0.1 M Tris / HCl buffer of pH 8.5 with 1.0 g of 5- (2-methylpropyl) hydantoin and incubated at 30 ° C. for 24 hours. The cell mass is then centrifuged off and 660 mg of L-leucine are isolated from the filtrate from the decomposition point 291 ° C. (from aqueous ethanol) [α] 20th D
Figure imgb0003
 = + 15.6 ° (20% aqueous hydrochloric acid).

Beispiel 6Example 6

  • a) 3,5 g feuchte Zellmasse von Nocardia spec. DSM 3306 - hergestellt nach Beispiel 5 - werden in 31,5 g einer 2 %igen wässrigen Lösung von Natriumalginat suspendiert und in 500 ml einer 0,1 M wässerigen lösung von Calziumchlorid-Dihydrat getropft.a) 3.5 g of moist cell mass from Nocardia spec. DSM 3306 - produced according to Example 5 - are suspended in 31.5 g of a 2% aqueous solution of sodium alginate and added dropwise in 500 ml of a 0.1 M aqueous solution of calcium chloride dihydrate.
  • b) 1,5 g des so erhaltenen Immobilisats werden in 10 ml 0,1 M Tris/HCl-Puffer vom pH 8,5 suspendiert mit 10 mg 5(2-Methylpropyl)-hydantoin versetzt und 18 Stunden lang bei 30° C incubiert. Durch Bestimmung mit L-Aminosäure-Oxidase wird ermittelt, daß 7,8 mg L-Leucin gebildet werden.b) 1.5 g of the immobilizate thus obtained are suspended in 10 ml of 0.1 M Tris / HCl buffer of pH 8.5 with 10 mg of 5 (2-methylpropyl) hydantoin and incubated at 30 ° C. for 18 hours . It is determined by determination with L-amino acid oxidase that 7.8 mg of L-leucine are formed.
Beispiel 7Example 7

Unter den Bedingungen des Beispiels 6 b werden aus 10 mg 5-(1-Methylpropyl)-hydantoin 3,2 mg L-Isoleucin gebildet.3.2 mg L-isoleucine are formed from 10 mg 5- (1-methylpropyl) hydantoin under the conditions of Example 6b.

Beispiel 8Example 8

Unter den Bedingungen des Beispiels 6 b werden aus 10 mg 5-(1-Methyl-ethyl)-hydantoin 2,1 mg L-Valin gebildet.2.1 mg of L-valine are formed from 10 mg of 5- (1-methyl-ethyl) -hydantoin under the conditions of Example 6b.

Beispiel 9Example 9

Unter den Bedingungen des Beispiels 6 b werden aus 10 mg 5,5'-Dithiobismethylen-bis-hydantoin 4,1 mg Cystin gebildet.4.1 mg of cystine are formed from 10 mg of 5,5'-dithiobismethylene-bis-hydantoin under the conditions of Example 6b.

Claims (3)

  1. Process for the preparation of L-amino acids of the general formula I
    Figure imgb0012
     wherein
    R₁ represents an alkyl radical having a maximum of 12 carbon atoms that is optionally substituted by hydroxy groups, mercapto groups, halogen atoms, amino groups, carbonyl groups or guanidino groups and/or optionally interrupted by oxygen atoms, nitrogen atoms or sulphur atoms, and, in the case of the mercapto compounds of the formula I, also the dithio compounds thereof, characterised in that the micro-organism Nocardia spec. DSM 3306 is allowed to act on a D,L-imidazolidinedione derivative of the general formula II
    Figure imgb0013
     wherein
    R₁ is as defined above or, in the case of the mercapto compounds of the formula II, alternatively on the dithio compounds thereof.
  2. Process for the preparation of L-amino acids of the general formula Ia
    Figure imgb0014
     wherein
    R₂ represents an alkyl radical having a maximum of 6 carbon atoms that is optionally substituted by a hydroxy group, a mercapto group, a methylthio group, an amino group, a carboxyl group or a guanidino group, and, in the case of the mercapto compounds of the formula Ia, also the dithio compounds thereof, characterised in that the micro-organism Nocardia spec. DSM 3306 is allowed to act on a D,L-imidazolidinedione derivative of the general formula IIa
    Figure imgb0015
     wherein
    R₂ is as defined above or, in the case of the mercapto compounds of the formula II, alternatively on the dithio compounds thereof.
  3. A process for the preparation of L-amino acids of the general formula Ib
    Figure imgb0016
     wherein
    R₃ represents an alkyl group having a maximum of 6 carbon atoms, a mercaptomethyl group or the grouping of the formula III
    Figure imgb0017
     characterised in that the micro-organism Nocardia spec. DSM 3306 is allowed to act on a D,L-imidazolidinedione derivative of the general formula IIb
    Figure imgb0018
     wherein
    R₄ represents an alkyl group having a maximum of 6 carbon atoms, a mercaptomethyl group or the grouping of the formula IV
    Figure imgb0019
EP88901022A 1987-01-23 1988-01-22 Process for producing l-aminoacids Expired - Lifetime EP0299028B1 (en)

Priority Applications (1)

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AT88901022T ATE83503T1 (en) 1987-01-23 1988-01-22 PROCESS FOR THE PRODUCTION OF L-AMINO ACIDS.

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE19873702384 DE3702384A1 (en) 1987-01-23 1987-01-23 METHOD FOR PRODUCING L-AMINO ACIDS
DE3702384 1987-01-23

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EP0299028A1 EP0299028A1 (en) 1989-01-18
EP0299028B1 true EP0299028B1 (en) 1992-12-16

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EP (1) EP0299028B1 (en)
JP (1) JPH01501841A (en)
AT (1) ATE83503T1 (en)
DE (2) DE3702384A1 (en)
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WO (1) WO1988005468A1 (en)

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GB2217319A (en) * 1988-04-19 1989-10-25 Synpharm Ltd Racemic and optically active fatty amino acids, their homo- abd hetero-oligomers and conjugates, the process of their production, their pharmaceutical composi
DE10115000C2 (en) * 2001-03-26 2003-02-20 Degussa Process for the preparation of enantiomerically enriched alpha-substituted carboxylic acids

Citations (1)

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Publication number Priority date Publication date Assignee Title
GB204531A (en) * 1922-10-02 1923-10-04 David Griffiths Improvements in and relating to automatic timing apparatus for cutting off the supply of the source of heat at the expiry of predetermined intervals

Family Cites Families (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4094741A (en) * 1976-02-04 1978-06-13 Kanegafuchi Kagaku Kogyo Kabushiki Kaisha Process for preparing D-(-)-N-carbamoyl-2-(phenyl or substituted phenyl)glycines
JPS5391189A (en) * 1976-12-30 1978-08-10 Kanegafuchi Chem Ind Co Ltd Preparation of d-n-carbamoyl-alpha-amino acids
US4211840A (en) * 1977-06-08 1980-07-08 Ajinomoto Company, Incorporated Method for producing D-α-amino acid
JPS557001A (en) * 1978-03-15 1980-01-18 Kanegafuchi Chem Ind Co Ltd Preparation of n-carbamoyl-d-thienylglycine
JPS55104890A (en) * 1979-02-06 1980-08-11 Kanegafuchi Chem Ind Co Ltd Production of d-alpha-aminoacids
JPS61177991A (en) * 1985-02-04 1986-08-09 Mitsui Toatsu Chem Inc Production of d-alpha-amino acid
JPS61181391A (en) * 1985-02-08 1986-08-14 Mitsui Toatsu Chem Inc Production of d-alpha-amino acid
JPS61285995A (en) * 1985-06-13 1986-12-16 Denki Kagaku Kogyo Kk Production of l-amino acid
JPH0659227B2 (en) * 1985-11-08 1994-08-10 三井東圧化学株式会社 Method for producing D-α-amino acid
JPS63112990A (en) * 1986-10-31 1988-05-18 Mitsui Toatsu Chem Inc Production of d-alpha-amino acid
US5259966A (en) * 1992-11-10 1993-11-09 The Lubrizol Corporation Low chlorine overbased calcium salts
JPH0823994A (en) * 1994-07-19 1996-01-30 Mitsubishi Heavy Ind Ltd Production of stable isotope-klabeled cytochrome c3 derived from sulfate reducing bacteria

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB204531A (en) * 1922-10-02 1923-10-04 David Griffiths Improvements in and relating to automatic timing apparatus for cutting off the supply of the source of heat at the expiry of predetermined intervals

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
Chemical Abstracts, vol. 107, No. 1, July 1987, (Columbus, Ohio, US), see pages 538-539, Abstract 5729g, & JP, A, 6200271 (DAIICHI KAGAKU YAKUHIN K.K.) 6 January 1987 *
Chemical Abstracts, vol. 93, No. 5, 4 August 1980, (Columbus, Ohio, US), see page 713, abstract 43950x, & JP, A, 8023994 (AJINOMOTO CO., INC.) 20 February 1980 *

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DE3876703D1 (en) 1993-01-28
JPH01501841A (en) 1989-06-29
FI884177A (en) 1988-09-12
FI884177A0 (en) 1988-09-12
ATE83503T1 (en) 1993-01-15
DE3702384A1 (en) 1988-08-04
US5071752A (en) 1991-12-10
WO1988005468A1 (en) 1988-07-28

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